Heegaard, Peter M. H.

Højbøge, Tina Rødgaard

Skovgaard, Kerstin

Stagsted, Jan

Online Research Database In Technology

                       General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.  • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal   If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.     Downloaded from orbit.dtu.dk on: Dec 19, 2017Innate immune responses to obesity in cloned and normal outbred domestic pigHøjbøge, Tina Rødgaard; Skovgaard, Kerstin; Stagsted, Jan; Heegaard, Peter Mikael HelwegPublication date:2012Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Højbøge, T. R., Skovgaard, K., Stagsted, J., & Heegaard, P. M. H. (2012). Innate immune responses to obesityin cloned and normal outbred domestic pig. Poster session presented at DFSTM Annual Meeting, Copenhagen,Denmark.Results and conclusion The variation in gene expression was found to be similar for the clones and the controls (not shown) and only a limited number of genes were affected by cloning (fig.1 and 2). In the two adipose tissues from the abdominal region (abdominal SAT and VAT(fig.2B and C)) 5 out of 6 significantly differentially expressed genes were down-regulated in the lean clones, whereas most differently expressed genes in both liver (fig.2A) and neck SAT (fig.2D) were up-regulated (6 out of 7). The same pattern was observed in the obese clones, although the number of genes affected was lower.  Systemically, circulating acute phase proteins (HP, CRP and ORM1) had increased serum concentrations in obese but not in lean clones, however this was only statistically significant for ORM1 (fig. 3). In conclusion, subtle but significant changes were identified in specific subsets of innate immune genes, especially in the lean clones, and this together with the finding that cloned pigs did not show lower inter-individual biological variation reduces the incentive to use cloned pigs for the study of innate immune gene regulation in obesity.  Materials and methods  All experimental procedures involving animals were approved by the Danish Animal Experimental Committee. Cloning was performed using somatic cell nuclear transfer as described in [3]. Donor cells were from cultured ear fibroblasts obtained from a Danish Landrace x Yorkshire (65%:35%) sow. Normal litters were kept as controls and were obtained after standard artificial insemination. Pigs were nursed by surrogate sows and weaned after 28 days. For an additional 2 months they were kept on standard diet after which they were individually housed. The pigs were then fed high-energy diet (containing 10% sugar and 10% soy oil) either ad libitum or 60% of ad libitum consumption. The pigs were killed at 9 months of age and blood and tissue samples were collected. A phenol/chloroform-method was used to extract mRNA from the tissues. qPCR was performed in 48.48 chips (FluiDigm) on a BioMark real-time PCR instrument (FluiDigm). Data was log2 transformed prior to t-test, F-test and ANOVA. ELISAs were performed to measure circulating concentrations of selected acute phase proteins.  Objective To evaluate the effect of cloning on the innate immune response in normal outbred pigs fed a restricted diet (60% of ad libitum diet) (lean) (n=9) compared to cloned pigs fed an ad libitum diet (obese) (n=9) or a restricted diet (lean) (n=8).   Introduction Pigs are widely used as a biomedical model for energy metabolism, obesity and inflammation in humans because of the similarities of porcine physiology and metabolic features to humans.  In theory the use of cloned animals should reduce biological variation. Therefore cloning may improve the applicability of pigs as a model for human obesity-induced inflammation.  However, previous findings have suggested that cloning may affect the phenotypes (e.g. [1]) including parts of the innate immune system [2].  Figure 1: Expression of measured genes in liver, abdominal SAT, VAT and neck SAT in clones and control pigs. Control Lean is set to 1. ODL=Over Detection Limit. NQ=Not Quantifiable.  Figure 3: Serum concentrations of ORM1, HP, CRP and IL-6 in controls (n=8, except for ORM 1where n=9) and clones (n=7, except for HP where n=8) measured with ELISAs. SEM is shown as error bars. *=P<0.05. Figure 2: Expression of significant genes in liver (A), subcutaneous fat (SAT) from the abdomen (B), intra-abdominal visceral adipose tissue (VAT) (C) and neck SAT (D) compared to control lean (set to 1) measured with qPCR. Control lean n=9, Clone lean n=8 (except for neck SAT where n=7) and Clone obese n=9. SEM is shown as error bars. *=P<0.05, **=P<0.01 and ***=P<0.001.   Innate immune responses to obesity in cloned and normal outbred domestic pig  Tina Rødgaard1, Kerstin Skovgaard1, Jan Stagsted2, Peter M. H. Heegaard1 1: Innate Immunology Group, National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark 2: Department of Food Science – Milk and Egg Science, Aarhus University, Tjele, Denmark References 1.  Clausen M, Christensen K, Hedemann M, et al. Metabolomic phenotyping of a cloned pig model.  BMC Physiology 2011;11:14. 2.  Carroll JA, Carter DB, Korte SW, Prather RS. Evaluation of the acute phase response in cloned  pigs following a lipopolysaccharide challenge. Domestic Animal Endocrinology 2005;29:564-72. 3. Kragh, P. M., Vajta, G., Corydon, T. J., Purup, S., Bolund, L., and Callesen, H. Production of  transgenic porcine blastocysts by hand-made cloning. Reproduction Fertility and Development 16,  315-18. 2004.    ORM1Control LeanClone LeanClone Obese0100200300400 *g/mL ORM1 in serumHPControl LeanClone LeanClone Obese0100020003000g/mL HP in serumCRPControl LeanClone LeanClone Obese0510152025g/mL CRP in serumIL-6Control LeanClone LeanClone Obese0.00.51.01.5ng/mL IL-6 in serumControl LeanClone LeanClone ObeseControl LeanClone LeanClone ObeseControl LeanClone LeanClone ObeseControl LeanClone LeanClone ObeseAPOA1 1 1,05 0,82 1 1,23 1,34 1 1,38 1,09 1 1,35 1,01CD14 1 0,96 0,88 1 0,76 0,82 1 0,91 1,09 1 0,80 0,85CD40 1 1,04 1,03 1 0,76 0,71 1 0,62 0,92 1 1,26 1,15C3 1 1,02 0,88 1 0,41 0,43 1 0,49 0,30 1 0,95 0,66CRP 1 1,67 1,27 1 1,06 1,36 1 0,59 0,53 1 0,73 0,90PDB-2 1 0,92 0,80 1 0,64 0,49 1 0,91 0,99 1 2,62 0,90FIB 1 1,18 0,93 ODL ODL ODL ODL ODL ODL ODL ODL ODLHP 1 1,28 0,96 1 0,23 0,42 1 0,20 0,13 1 0,92 0,79IL-8(a) NQ NQ NQ 1 0,58 0,45 1 0,44 0,21 1 3,98 1,47IL-1B 1 0,89 0,66 1 1,65 0,96 1 0,80 0,83 1 2,07 1,55IL-6 NQ NQ NQ 1 2,04 1,34 1 0,73 0,74 1 1,19 0,43IL-8(b) 1 0,92 1,46 1 0,57 0,46 1 0,49 0,23 1 3,11 1,62IL-10(a) 1 1,36 1,21 1 1,13 0,81 1 1,17 1,20 1 2,03 1,26pigMAP/ITIH4(b) 1 1,61 1,27 1 1,46 1,34 1 1,21 0,86 1 1,25 0,98pigMAP/ITIH4(a) 1 1,71 1,34 1 1,44 1,61 1 0,87 0,48 1 1,48 1,03SAA 1 2,59 2,43 1 0,07 0,23 1 0,37 0,38 1 1,27 1,65TGFB1 1 0,88 0,89 1 0,82 0,87 1 0,75 0,80 1 1,03 1,17TNF-α 1 1,25 0,90 1 1,58 1,19 1 1,02 1,52 1 1,61 1,33TLR4 1 0,87 1,04 1 1,15 0,77 1 1,17 1,07 1 1,05 0,85LBP 1 1,46 1,18 NQ NQ NQ NQ NQ NQ NQ NQ NQAOAH 1 1,17 1,16 1 1,29 1,25 1 1,35 1,52 1 1,66 1,53CXCL10 1 2,72 1,29 1 1,99 0,71 1 3,04 0,73 1 5,88 0,79CCL3LI 1 1,11 0,75 1 1,22 0,96 1 0,76 0,64 1 0,96 1,09CCL2 1 1,06 1,98 1 0,84 0,66 1 0,60 0,51 1 1,10 1,01CCL5 1 0,99 1,20 1 1,01 1,07 1 1,50 0,77 1 1,40 1,12PAFA-H1B1 1 0,91 0,92 1 0,87 0,92 1 0,96 1,08 1 0,76 0,91SFTPA1 1 2,16 3,59 NQ NQ NQ NQ NQ NQ NQ NQ NQIL-1RN 1 0,99 0,86 NQ NQ NQ NQ NQ NQ NQ NQ NQNFKB1 1 0,93 1,07 1 0,83 0,90 1 0,84 0,92 1 0,95 0,92NFKBIA 1 1,18 0,93 1 1,28 1,05 1 0,97 0,70 1 2,09 1,01ORM1 1 1,10 1,06 ODL ODL ODL ODL ODL ODL ODL ODL ODLTF 1 1,11 0,99 1 0,49 0,16 1 0,11 0,07 1 1,54 0,86CD36 1 0,99 0,83 1 1,24 1,49 1 1,26 1,18 1 0,66 0,86COX-2 1 1,63 4,79 1 1,63 1,02 1 1,33 0,46 1 2,66 0,65IL-10(b) 1 1,71 1,11 NQ NQ NQ NQ NQ NQ NQ NQ NQCD200 1 0,93 0,93 1 0,72 0,59 1 0,97 0,98 1 0,85 0,79IL-18 1 0,90 1,33 1 0,67 0,71 1 0,69 1,21 1 2,13 1,93IFNG 1 0,88 0,96 NQ NQ NQ NQ NQ NQ NQ NQ NQTNFAIP3 1 1,02 0,93 1 1,21 0,97 1 1,14 1,14 1 1,14 0,83CD163 1 0,94 1,32 1 1,22 1,21 1 1,16 1,35 1 1,53 1,380 1 <5LiverAbdominal SAT VAT Neck SATLiverSAASFTPA1pigMAP/ITIH4IL-8012345 Control LeanClone Lean***Clone Obese*Relative quantitiesAbdominal SATC3HPSAA TNF-0.00.51.01.52.02.5Control LeanClone Lean*** *Clone ObeseRelative quantitiesVATC3SAAIL-8pigMAP/ITIH4CCL5COX-20.00.51.01.52.02.5Control LeanClone Lean** ***Clone Obese*** *Relative quantitiesNeck SATIL-10NFKBIAAOAHCD360123 Control LeanClone Lean* ***Clone Obese*Relative quantitiesA BC DTina Rødgaard (tinro@vet.dtu.dk) Innate Immunology Group National Veterinary Institute Technical University of Denmark Bülowsvej 27, Building B, room B7 1870 Frederiksberg C Denmark   

Innate immune responses to obesity in cloned and normal outbred domestic pig

http://orbit.dtu.dk/ws/files/60237747/Poster_til_WIRM_marts_2012.pdf

Innate immune responses to obesity in cloned and normal outbred domestic pig

Abstract

Similar works

Full text

Available Versions

Online Research Database In Technology